【作者】 张健；

【导师】 任炳忠；

【作者基本信息】 东北师范大学 ， 动物学， 2011， 博士

【摘要】 天牛科Cerambycidae是鞘翅目Coleoptera中最大的科之一,全世界约有45000种,是鞘翅目昆虫中形态变异最具多样性的类群之一。到2005年止,中国天牛记录已有3100余种。天牛科昆虫绝大多数取食植物,部分种类的天牛是林木的重要害虫,严重危害果树和观赏类树木;其它种类的天牛危害落叶树和阔叶树,少部分的天牛危害农作物。大多数种类的天牛在自然生态系统中具有重要的生态价值。由于天牛与其寄主植物紧密的协同进化,天牛可以作为森林健康和多样性的重要生物指示信号。本研究共整理鉴定出吉林省天牛科昆虫7亚科102属223种,并对吉林省天牛科昆虫的区系组成及分布特点进行了分析。在所整理鉴定的223种天牛中,各分布区的种类由多至少依次为东北区222种,华北区111种,蒙新区108种,华中区79种,华南区60种,西南区42种,青藏区16种。各亚科所占比例分别为沟胫天牛亚科35属84种,占总种数的37.67%;天牛亚科26属55种,占总种数24.66%;锯天牛亚科4属4种,占总种数1.79%;花天牛亚科32属67种,占总种数30.04%;幽天牛亚科3属6种,占总种数2.69%;椎天牛亚科1属1种,占总种数0.45%;膜花天牛亚科1属6种,占总种数2.69%。自从天牛科这一分类单元建立以来,对于天牛科亚科之间的系统发育关系这一问题国内外许多学者提出个许多不同的见解,至今尚无定论。本文主要就吉林省分布数量较多的4亚科天牛间的系统发育关系进行探讨,试图解决这4个亚科与其它亚科的相互关系,并对传统分类学提出的观点的可靠性进行验证。本研究共采用4亚科31属36种天牛进行研究。1生物化学方面:采用聚丙烯酰胺凝胶电泳技术对3亚科9属11种天牛的酯酶同工酶进行研究,探讨酯酶同工酶技术应用于天牛亚科级阶元分类的有效性。结论如下:(1)从酶带数量上来看,沟胫天牛亚科相对数量较多,9-13条不等;花天牛亚科酶带数量较少,2-6条不等,多为3条。从酶活性来看,沟胫天牛亚科类群酶活性强于其它两个亚科。(2)聚类分析结果表明,根据酶带的迁移率,3亚科昆虫有效地归为3个分支,其中天牛亚科和花天牛亚科亲缘关系较近,聚为一个分支,后与沟胫天牛亚科相聚。在标本数量足够多的情况下,利用酯酶同工酶分析天牛亚科之间的亲缘关系是可行的。但在解决近缘属种的关系问题上,酯酶同工酶存在一定的局限性。(3)由于所选取的分类单元数量较少,对于天牛科亚科间关系这一系统发育问题,还需结合形态学手段及分子生物学手段得到的结果联合起来分析,才能得到更为可靠的结果。2分子生物学方面:提取了天牛科4亚科31属36种昆虫的总DNA。采用3对通用引物扩增了2段线粒体基因部分序列和1段核基因部分序列,获得长度为771bp的COI基因序列36条,长度为501bp的16SrDNA基因序列36条,长度为772bp的28SrDNA基因序列32条;采用Clustal v1.8、Bioedit v7.0.9和MEGA 4.0软件对DNA序列的碱基组成、碱基替换及遗传距离进行分析;基于28SrDNA基因序列和3段基因联合数据集采用PAUP 4.0的MP法、NJ法、PHYML v2.4.4的ML法以及MrBayes 3.1.2的贝叶斯法分别重建系统发育树,研究得到以下结果:(1)共向GenBank数据库提交新序列104条,涉及天牛科4亚科31属36种,包含3种基因。(2)在天牛科内线粒体基因序列为“AT”碱基丰富序列,线粒体基因序列的颠换高于转换,而核基因序列的转换稍高于颠换。(3)本论文基于28SrDNA基因序列的建树结果有效的将锯天牛亚科、花天牛亚科、沟胫天牛及天牛亚科聚在不同的分支,从分子角度证明了4亚科的单系性。(4)本论文基于联合基因系统发育树的结果与王乔和蒋书楠(1991)以植物学和古地理学为依据,结合形态特征和天牛生物学特性讨论天牛科主要亚科的演化关系部分结果一致。认为沟胫天牛亚科和天牛亚科是较为原始的类群,锯天牛亚科和花天牛亚科是较为进化的类群。3触角感器显微结构方面:本论文结果表明,曲纹花天牛和橡黑花天牛雄性锥形感器2的数量显著多于雌性。在其它类群的昆虫中,也发现同种昆虫雌雄个体感器数量存在差异的现象。同属的两种天牛,不同类型感器在数量、分布上存在差异。目前,由于不同学者对昆虫触角感器的分类和命名采用的标准不同,不同描述也存在着较大的差异。触角感器作为分类特征和系统发育研究的依据仍存在一定的局限性。此外,有研究发现,昆虫感器的数量并不是固定不变的,例如蝗虫,会因发育阶段不同,生理状态不同而改变。目前国内外学者对昆虫触角感器的研究大多集中于单一种类昆虫感器的类型、数量及分布上,并没有对触角感器作为昆虫分类研究和系统发育研究的可靠性作出定论。本论文通过现代生物学技术对基于形态学角度的天牛科传统分类学观点进行了验证,现代生物学技术具有准确、客观、灵敏等优点,弥补了传统形态学方法的缺点和不足。DNA序列数据具有明确的遗传基础,其数据量的大小仅受基因的大小限制。形态数据的优点在于它可以从化石和保存的标本上得到,并可从个体发育角度进行解释。而且,有些问题只能通过形态特征来解决,另一些问题则要应用分子数据来解决。大量研究表明,将分子与形态结合起来比单一方法可对问题作出更好的描述和解释。更多还原

【Abstract】 Longicorn beetles (Coleoptera: Cerambycidae), with more than 45000 known species, comprised one of the largest and most diverse groups of Coleoptera. The total number of longicorn beetles recorded from China has exceeded 3100 species up to 2005. Most longicorn beetles feed on plants. Several species of cerambycid beetles are known to damage living trees, and may seriously affect orchards and ornamental trees; others may damage coniferous or hardwood lumber, and a few taxa can damage row crops. The majority of species occur within uncultivated or forested habitats and are of great ecological importance within these natural ecosystems. Because of their close evolutionary linkage to their host plants, cerambycid beetles may be valuable bio-indicators of forest health and diversity.223 species representing 64 genera of 7 subfamilies of Cerambycidae in Jilin Province were identified and compiled in this study. The faunal composition and distribution characters were analysed. The 223 species are distributed in 7 subregions. 222 speices in Northeast China subregion, 111 speices in North China subregion, 108 speices in Mongolia Xinjiang subregion, 79 speices in Middle China subregion, 60 speices in South China subregion, 42 speices in Southwest China subregion, and 16 speices in Qinghai Xizang subregion. 84 species of 35 genera of Lamiinae account for 37.67%; 55 species of 26 genera of Cerambycinae account for 24.66%; 4 species of 4 genera of Prioninae account for 1.79%; 67 species of 32 genera of Lepturinae account for 30.04%; 6 species of 3 genera of Aseminae account for 2.69%; 1 species of 1 genera of Spondylidinae account for 0.45%; 6 species of 1 genera of Necydalinae account for 2.69%.Although the phylogenetic hypotheses of the subfamilial relationships of Cerambycidae were put forward by many scholars since Cerambycidae was found, the higher classification of the Cerambycidae has not been sufficiently elucidated so far. The phylogenetic relationships of 4 subfamilies broadly distributed in Jilin Province were reconstructed in this study. We also tested the traditional classification views on Cerambycidae through modern bio-technology. 36 species of 31 genera of 4 subfamilies in Cerambycidae were analysed in this study.1 BiochemistryIn this part, the EST isoenzyme of 11 species longicorn beetles of 9 genera of 3 subfamilies were analyzed by using the technique of vertical slab poly-acrylamidae gel electrophoresis, and clustering analysis of these species was researched based on their relative mobilities to test if EST isoenzyme analyses can be feasible to resolve the phylogenetic relationships of Cerambycidae at the subfamily level. The results are as follows:(1) The number of the isoenzyme bands of Lamiinae ranged from 9 to 13 and the number of the isoenzyme bands of Lepturinae ranged from 9 to 13. The enzyme activity of Lamiinae was stronger the other two subfamilies.(2) The 11 species of 3 subfamilies were grouped in 3 clades based on relative mobilities by clustering analysis. The results indicated that it was feasible to reconstruct the phylogenetic relationships of Cerambycidae using this method when the taxon was enough. However, the clustering analysis based on their relative mobilities cannot solve the phylogenetic relationships at the genera level.(3) The clustering analysis results of Cerambycidae based on their relative mobilities of the isoenzyme bands were consistent with the results inferred from morphological classification at subfamily level. Because the number of species in Cerambycidae is so large, it is necessary to solve the phylogenetic relationships combined with different methods.2 Molecular BiologyTotal genomic DNA was extracted from 36 species of 31 genera of 4 subfamilies in Cerambycidae. Amplification of the 2 fragment mitochondrial genes (COI 771bp; 16SrDNA 501bp) and 1 fragment ribosomal gene (28SrDNA 772bp) was performed using 3 pairs of universal primer. The base composition, base substitution and genetic distance of DNA sequences were analysed using Clustal v1.8, Bioedit v7.0.9 and MEGA 4.0 software. The phylogenetics relationships were reconstructed from the nuclear 28S ribosomal DNA and combined data using PAUP4.0 (maximum parsimony method, neighbor joining method), PHYML v2.4.4 (maximum likelihood method) and MrBayes 3.1.2 (Bayesian inference method). The results are as follows:(1) We submitted 104 new DNA sequences of 3 genes to GenBank, covering 36 species of 25 genera of Cerambycidae.(2) The family Cerambycidae is“AT”base-bias in the DNA sequences of mitochondrial genes, and transversion is higher than transition, but transition is higher than transversion in ribosomal gene sequences.(3) The phylogenetic trees were inferred from the nuclear 28S ribosomal DNA. The species of the 4 subfamilies were divided into 4 clades. The monophyly of the 4 subfamilies was resolved respectively.(4) The phylogenetics relationships inferred from combined gene sequences in our study were partically congruent with the results of previous investigations based on botany, geography, morphological and biology characteristics of Cerambycidae by Wang and Jiang (1991). Lamiinae and Cerambycinae are the older groups and Prioninae and Lepturinae are the younger groups.3 Fine structure of antennal sensillaThe average number of sensilla basiconica type 2 of Leptura arcuata Panzer and Leptura aethiops Poda on the antennae of the males was significant greater than that on the antennae of the females. The differences of the number of antennal sensilla between males and females were also found in other insects. The number and distribution of antennal sensilla of both species of the longicorn beetles in one genera were different.Because different scholars classified the sensilla into various types based on different standard and the descripition of the same type of the sensilla bear differences. So it is cautious to apply the characters of sensilla to identify specimen and solve the phylogenetics relationships of insects.We tested the traditional classification views on Cerambycidae through modern bio-technology. Modern bio-technology bear the characters of accuracy, impersonality and delicacy and play as a remedy for the traditional classification. DNA sequences have explicit genetic foundation. The amount of the DNA data was controlled by the size of gene. The merits of morphological data lie in that these data could be collected from fossil and specimen. And morphological data can be explained from individual growth. Some questions only can be solved by morphological characters, and the other questions only can be solved by molecular data. A large amounts of research demonstrated that the results of phylogenetics analyses based on molecular and morphological connected data were more convinced compared with the sole analyses.更多还原